Nā huahana

Ke Kaʻina Metagenomic -NGS

ʻO ka Metagenome e pili ana i ka hōʻiliʻili ʻana o ka nui o nā mea moʻo o kahi kaiāulu hui o nā mea ola, e like me ka metagenome kaiapuni, metagenome kanaka, a me nā mea ʻē aʻe.ʻO ka metagenomic sequencing he mea hana molecular i hoʻohana ʻia e kālailai i nā mea genomic hui ʻia i unuhi ʻia mai nā laʻana kaiapuni, e hāʻawi ana i ka ʻike kikoʻī i ka ʻano like ʻole a me ka nui o nā ʻano, ke ʻano o ka heluna kanaka, ka pilina phylogenetic, nā genes hana a me ka pilina pili me nā mea kaiapuni.

Papahana:ʻO Illumina NovaSeq Platform

Kāhea iā mā˚ou

Nā kikoʻī lawelawe

Nā hopena hōʻike

Nānā Hiʻohiʻona

Pono lawelawe

● Hoʻokaʻawale a mahiʻai ʻole no ka hoʻopili ʻana i ke kaiāulu microbial

● Hoʻoholo kiʻekiʻe i ka ʻike ʻana i nā ʻano haʻahaʻa haʻahaʻa i nā laʻana kaiapuni

● Hoʻohui ka manaʻo o "meta-" i nā hiʻohiʻona olaola a pau ma ka pae hana, ka pae ʻano a me ka pae gene, e hōʻike ana i kahi hiʻohiʻona ikaika e pili kokoke ana i ka ʻoiaʻiʻo.

● Hoʻoulu ʻo BMK i ka ʻike nui ma nā ʻano laʻana like ʻole me ka ʻoi aku o 10,000 mau laʻana i hana ʻia.

Nā kikoʻī lawelawe

Papahana	Ke kaʻina ʻana	Manaʻo ʻikepili	Ka manawa huli
ʻO Illumina NovaSeq Platform	PE150	6 G/10 G/20 G	45 mau lā hana

Nā loiloi bioinformatics

● Ka mana o ka maikaʻi o ka ʻikepili maka

● Hui Metagenome

● ʻAʻole-redundant gene set and annotation

● Ka nānā 'ana i nā 'ano like 'ole

● Ke kālailai ʻokoʻa hana genetic

● Ka nānā ʻana ma waena o ka hui

● Kānāwai hui kūʻē i nā kumu hoʻokolohua

Nā Koina Laʻana a me ka Hoʻouna ʻana

Nā Koina Laʻana:

No ka meaNā unuhi DNA:

ʻAno Laʻana	Ka nui	Hoʻopaʻa	Maemae
Nā unuhi DNA	＞ 30 ng	＞ 1 ng/μl	OD260/280= 1.6-2.5

No nā laʻana kaiapuni:

ʻAno laʻana	Manaʻo ʻia ke kaʻina hana laʻana
lepo	Ka nui o ka laʻana: approx.5 g;Pono e wehe ʻia nā mea maloʻo i koe mai ka ʻili;E wili i nā ʻāpana nui a hele i ka kānana 2 mm;ʻO nā laʻana Aliquot i loko o ka sterile EP-tube a i ʻole cyrotube no ka mālama ʻana.
Feces	Ka nui o ka laʻana: approx.5 g;E hōʻiliʻili a hāʻawi i nā laʻana i loko o ka sterile EP-tube a i ʻole cryotube no ka mālama ʻana.
ʻO loko o ka ʻōpū	Pono e hana ʻia nā laʻana ma lalo o ke kūlana aseptic.Holoi i ka ʻiʻo i hōʻiliʻili ʻia me ka PBS;Centrifuge i ka PBS a ohi i ka precipitant ma EP-tubes.
Sludge	Ka nui o ka laʻana: approx.5 g;E hōʻiliʻili a hāʻawi i kahi laʻana o ka lepo i loko o ka EP-tube sterile a i ʻole cryotube no ka mālama ʻana
Ke kino wai	No ka laʻana me ka liʻiliʻi o ka microbial, e like me ka wai paipu, ka wai punawai, a me nā mea ʻē aʻe, E hōʻiliʻili i ka liʻiliʻi o 1 L wai a hele i ka kānana 0.22 μm e hoʻonui i ka microbial ma ka membrane.E mālama i ka membrane i loko o ka paipu sterile.
ʻili	E ʻoki pono i ka ʻili o ka ʻili me ka pulupulu maʻemaʻe a i ʻole ka lāʻau lapaʻau a waiho i loko o ka paipu sterile.

Manaʻo ʻia ka hāʻawi laʻana

E hoʻokuʻu i nā laʻana i loko o ka hau hau no 3-4 mau hola a mālama i loko o ka wai nitrogen a i ʻole -80 degere a hiki i ka mālama lōʻihi.Pono ka hoʻouna laʻana me ka hau maloʻo.

Holo Hana Hana

Hāʻawi laʻana

Kūkulu hale waihona puke

Ke kaʻina ʻana

ʻIkepili ʻikepili

Nā lawelawe ma hope o ke kūʻai aku

Mua: ʻO ke Kaʻina Metagenomic-Nanopore

Aʻe: ʻO Chromatin Immunoprecipitation Sequencing (ChIP-seq)

1.Histogram: Hoʻolaha ʻano ʻano

2.Functional genes annotated to KEGG metabolic pathways

3. Heat map: Nā hana like ʻole e pili ana i ka nui o nā gene pili4.Circos o CARD antibiotic kū'ē genes

Hihia BMK

ʻO ka laha ʻana o nā genes pale antibiotic a me nā maʻi bacterial ma ka hoʻomau ʻana o ka lepo-mangrove root continuum

Paʻi ʻia:Nūpepa o nā Mea Pilikino, 2021

Hoʻolālā kaʻina hana:

Mea waiwai: Nā ʻāpana DNA o nā ʻāpana ʻehā o nā kumu lāʻau mangrove e pili ana: lepo kanu ʻole, rhizosphere, episphere a me nā keʻena endosphere.
Papahana: Illumina HiSeq 2500
Nā pahuhopu: Metagenome
16S rRNA gene V3-V4 māhele

Nā hopena koʻikoʻi

Ua hoʻoponopono ʻia ka metagenomic sequencing a me ka metabarcoding profiling ma ka ʻaʻa lepo o nā kumu lāʻau mangrove i mea e aʻo ai i ka hoʻolaha ʻana o nā genes resistance antibiotic (ARGs) mai ka lepo i nā mea kanu.Ua hōʻike ʻia ka ʻikepili Metagenomic he 91.4% o nā genes resistance antibiotic i ʻike maʻamau i loko o nā keʻena lepo ʻehā i ʻōlelo ʻia ma luna, kahi i hōʻike ʻia i ke ʻano mau.ʻO 16S rRNA amplicon sequencing i hana i 29,285 sequencing, e hōʻike ana i 346 mau ʻano.I ka hui pū ʻana me ka hoʻopili ʻana i nā ʻano mea ma o ka amplicon sequencing, ua ʻike ʻia kēia hoʻolaha ʻana he kūʻokoʻa i ka microbiota pili i ke aʻa, akā naʻe, hiki ke hoʻomaʻamaʻa ʻia e ka mobile o nā mea genetic.Ua ʻike kēia haʻawina i ka kahe ʻana o nā ARG a me nā pathogens mai ka lepo i loko o nā mea kanu ma o ka hoʻomau ʻana o ka lepo-aʻa.

Kuhikuhi

Wang, C. , Hu, R. , Strong, PJ , Zhuang, W. , & Shu, L. .(2020).ʻO ka laha ʻana o nā genes kūpaʻa antibiotic a me nā maʻi bacterial ma ka lepo-aʻa kumu mangrove.Nūpepa o nā mea pōʻino, 408, 124985.